EP2165193B1 - Dispositif de surveillance de la présence de germes microbiens dans l'eau - Google Patents

Dispositif de surveillance de la présence de germes microbiens dans l'eau Download PDF

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Publication number
EP2165193B1
EP2165193B1 EP08756847.3A EP08756847A EP2165193B1 EP 2165193 B1 EP2165193 B1 EP 2165193B1 EP 08756847 A EP08756847 A EP 08756847A EP 2165193 B1 EP2165193 B1 EP 2165193B1
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Prior art keywords
reactor
water
filter
chamber
pump
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German (de)
English (en)
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EP2165193A1 (fr
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Thomas Lendenfeld
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Microlan BV
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Microlan BV
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
    • C12M33/14—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus with filters, sieves or membranes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
    • C12M41/38—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of metabolites or enzymes in the cells
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor

Definitions

  • the invention relates to a device for continuous, periodic monitoring of water for microbial germs, comprising a reactor, at least one water supply for the water to be tested in the reactor, at least one water discharge from the reactor chamber, at least one reagent feed into the reactor and at least one Metering device for reagents, at least one measuring device for detecting the presence of microorganisms and / or their metabolic products, with pumps / valve means in the inlet / outlet lines of the reactor and with a controller for programmed control of at least the pump / valve means.
  • water is understood in the context of the invention, any type of liquid whose main component is water, for example, drinking water, mineral water, water used for drinking water, river and lake water, technical process water, eg cooling water, circulating water, biotechnological process solutions, eg Fermentation solutions, wastewater and purified waste water, etc., and food whose main constituent is water, eg milk and milk products or products of the beverage industry.
  • the mentioned problem is also the document DE 86 23 413 U1 refer to.
  • This document discloses a measuring device for detecting coliform bacteria in water samples in which hydrogen produced by the bacteria is detected.
  • the meter uses one or more sealed incubation containers into which the water to be tested, a nutrient medium and for cleaning a disinfectant (chlorine water) can be introduced.
  • the starting point of this known measuring device is the view that a preconcentration of bacteria for many reasons is not desirable, in particular because filters clogged and the bacteria concentration on the filter could be too high and also the preconcentration on the filter membrane was irreversible. This lead to the fact that the use of a filter membrane did not allow a simple automation.
  • the sample to be examined is pumped by means of a pump via an intermediate container, a three-way valve and another pump in a mixing chamber and with the aid of valves and a pump reagents are pumped into the mixing chamber.
  • the sample enters an incubator with a capillary.
  • the sample is transferred to another mixing chamber, where a reagent is added again via a valve and a pump.
  • the sample After passing through another capillary, the sample enters a measuring cell, where it is measured by fluorescence optics.
  • the described device can not achieve the required sensitivity in order to meet the drinking water requirements; on the other hand, the consumption of reagents due to the continuous mode of operation is relatively high, which means that considerable costs can be expected during operation.
  • WO 2005/083109 A1 describes a method for measuring contaminants in a medium by passing a certain volume of the medium through a filter, then contacting the inflow side of the filter with a substrate that produces a detectable component in interaction with a contaminant ; this document does not disclose a device that performs the measuring method.
  • US 2007/0003997 A1 describes a method and apparatus for measuring bacteria in a fluid sample.
  • samples are still taken and taken to a laboratory, where any existing germs are detected after incubation on a nutrient medium.
  • the reactor has a reactor chamber and a filtrate separated by this filter, the at least one water supply and the at least one reagent feed into the reactor chamber and the at least one water drainage the filtrate chamber leads, and the controller is adapted by means of the pump / valve means a predetermined amount of water in the reactor chamber and pass through the filter and introduce a predetermined amount of reagent into the reactor chamber, the controller is adapted to reverse the flow of water through the reactor by means of the pump / valve means and the filtrate chamber is above the reactor chamber.
  • a device which allows automated measurement over long periods of time and thanks to its design is versatile, so that, for example, no restriction on certain germs, detection methods or quality of the water to be examined is given by the reversal of the flow of water through the Reactor can be measured on the one hand in reflux, whereby the dilution occurring in some cases by the passage of the filter and thus a signal reduction does not occur, and on the other hand, the cleaning of the apparatus can be facilitated.
  • the measuring device In an embodiment which can be realized in a particularly favorable manner, provision is made for the measuring device to be set up for the measurement downstream of the filtrate chamber.
  • a concentration unit is arranged in front of the measuring device. It has proven to be particularly effective when the concentration unit operates on the principle of chromatographic separation.
  • the flexibility of the device can be increased by arranging the measuring device for measurement in the reactor chamber and / or for measurement in the filtrate chamber.
  • the controller is set up to circulate the flow of water through the reactor by means of the pumps / valve means.
  • the controller is set up to detect and / or prescribe the measuring volume conveyed via the pump.
  • a filter replacement plate is provided with at least two filter inserts, which, sealed against the reactor housing, is displaced so that each filter insert as an effective filter reactor and filtrate separates or that the filter replacement plate one of the Control is assigned to driven drive.
  • the filter is substantially planar and inclined to the horizontal so that gas bubbles in the reactor chamber can stroke the filter surface upwards. It is particularly advantageous if the filtrate chamber discharge openings for draining the water into a first and a second drainage, wherein the discharge opening of the first water discharge at the highest point of the filtrate and the discharge opening of the second water discharge, which supplies the water to the measuring device in a is arranged substantially normal to the filter plane extending symmetry axis of the reactor.
  • a vent line and a vent valve may be provided for removing air or gas bubbles from the reactor at least for the reactor chamber. If a filter exchange plate is present, the solution offers the fact that the filter exchange plate has a vent recess which releases in a defined displacement position of the plate a connection between the reactor and / or filtrate chamber and an outwardly lead the vent line.
  • Fig. 1 the water to be examined - for the definition of "water” see above - via a water supply line 1, a first 3-way valve 2, a pump 3 and a second 3-way valve 4 to a reactor 5 and from there via a third 3-way valve 6, a fourth 3-way valve 7 and a fifth 3-way valve 8 via a water outlet 9 again derived.
  • the reactor 5 has a reactor chamber 11 and a, here overlying, separated by a filter 12 Filtrathunt 13.
  • the temperature within the reactor can be detected by means of a temperature sensor 14 and a corresponding information to the controller 10th to get redirected.
  • the reactor 5 can be heated by means of a heater 15, which is controlled via the controller, expediently starting from the information of the temperature sensor 14.
  • a measuring device 16 is arranged which communicates with a peripheral unit 17 for this measuring device, on the other hand the latter being able to communicate with the controller 10.
  • Reagents from storage containers 20a, 20b, 20c and 20d can be introduced into the reactor chamber 11 via reagent feed lines 18a, 18b, 18c and 18d and metering valves 19a, 19b, 19c and 19d which can be controlled by the controller 10.
  • a pressure measuring device 21 is arranged, which forwards the value of the current pressure to the controller 10, so that this, for example via the delivery rate of the pump 3, can maintain the pressure at a predeterminable value ,
  • a transverse line 26 connects the output of the filter chamber 13 with the second 3-way valve 4, a transverse line 27, the input of the reactor chamber 11 with the fourth 3-way valve. 7
  • FIG Fig. 2 Before discussing the function or the various operating modes of a device according to the invention, further details with reference to FIG Fig. 2 explained, in which like reference numerals for like parts as in Fig.1 be used and already in Fig. 1 explained parts are omitted.
  • the Fig. 2 shows a reactor 5d, which is inclined obliquely with respect to the horizontal. Due to the inclination of the reactor 5d and the filter plate 31 is inclined.
  • a magnetic stirrer 28 is provided here which has a motor 29 and a stirring magnet 30 inside the chamber 11.
  • the motor 29 is in turn from the central Control 10 controlled.
  • Other technologies may be used to maintain movement in the reactor chamber 11, if desired.
  • the inclination of the reactor 5d results in a substantially horizontal area in the reactor chamber 11 for the stirring magnet 30.
  • the stirring magnet becomes 30 held in the lowest position due to the geometry of the reactor chamber 11 by gravity.
  • the apparatus has a filter replacement plate 31 with three filter cartridges 32a, 32b and 32c.
  • These filter cartridges may be, for example, commercially available ceramic filters.
  • a controlled by the controller 10 drive 33 can now move the filter replacement plate 31, which is sealed against the reactor housing, so that in each case another filter insert as an effective filter separates the reactor from the filter chamber In the position shown, the filter insert 32b is effective.
  • any air or gas bubbles present in the reactor chamber 11 can be swept upwards along the filter surface, pressed through the filter 12 into the filtrate chamber 13 and discharged.
  • two water outlets 9a and 9b lead from the filtrate chamber 13 of the reactor 5d.
  • the discharge opening 9aa of the first water discharge 9a is located substantially at the highest point of the filtrate chamber 13 and serves to drain the water from the reactor 5d during the filtration process.
  • any air bubbles which strike the reactor chamber upwards along the filter surface and pass through the filter 12 can escape via the first water outlet 9a.
  • the discharge port 9ba of the second water discharge 9b, through which the water is supplied to the meter 16, is disposed in the axis of symmetry L of the reactor 5d.
  • the symmetry axis L of the reactor 5d runs essentially normal to the filter surface.
  • This arrangement has the advantage that in the measuring operation in which the solution to be measured is guided from the reactor chamber 11 through the filter 12 into the filtrate chamber 13 and from there via the second water outlet 9b in the measuring device 16, no discrimination of the reaction solution - due to different Paths from the filter surface to the discharge port 9ba the second water discharge 9b - takes place and therefore mixing effects and associated signal reduction are kept small.
  • the water outlets 9a and 9b open via a 3-way valve 6 'in a water discharge.
  • vent line 34 and a vent valve 35 may be provided for the reactor chamber 11, so that venting can be performed if necessary or at certain times of the procedure. It is also useful here, if the usually just trained filter is inclined to the horizontal so that gas bubbles in the reactor chamber 11 stroke over the filter surface upwards and can get to the inlet of said vent line 34.
  • Fig. 3 shows the valve positions and the associated flow in the "Filtration" operating mode.
  • the sample is continuously sucked in via the water supply line 1 by means of the pump 3 and passed through the reactor chamber 11, the filter 12 and the filtrate chamber 13 to the discharge line 9.
  • the filtered volume of water sample is measured or metered.
  • the revolutions of the pump 3 can be detected, or a flow meter 21a can be used to detect the volume delivered by the pump 3.
  • This flow meter 21a provides corresponding information to the controller 10 and this can turn off the pump 3 or close valves after reaching a certain volume delivered. Microbial germs are retained on the filter 12.
  • the operating mode "Filtration” is followed by the operating mode "Dosing and incubation” according to Fig. 4
  • reagents from the storage containers 20a, 20b, 20c and 20d are initially introduced into the reactor chamber 11 via the reagent feed lines 18a, 18b, 18c and 18d and metering valves 19a, 19b, 19c and 19d which can be controlled by the controller 10
  • the displaced volume leaves the reactor via the filter, so that the volume in the reactor chamber remains constant.
  • the metered addition of the reagents can optionally be done at different times of incubation.
  • the heater 15 controlled via the temperature sensor 14 and the controller 10 as well as the magnetic stirrer 28 ensure optimum reaction conditions
  • the subsequent measuring step takes place in the operating mode "Measurement" Fig. 5 ,
  • the pump 3 is turned on again and water flowing through the supply line 1 presses the measuring solution located in the reactor chamber 11 through the filter 12 via the filtrate chamber 13 and from here via the third 3-way valve 6 into a measuring cell of the measuring device 16
  • Peripheral unit 17 and / or the control 10 outputable measured value describes the concentration of the analytes in the reactor chamber at the end of the incubation. variants
  • the measurement options and measurement locations are related below Fig. 7 described.
  • E. coli Escherichia coli
  • E. coli is an indicator of fecal contamination and is routinely routinely monitored worldwide. used by drinking water.
  • a defined volume of sample is filtered by the pump via the filter in the reactor chamber.
  • Buffer and substrate solution are added to this volume of the reactor chamber containing the enriched bacteria.
  • the buffer solution provides ideal conditions for the enzymatic reaction
  • the substrate solution contains a substrate which can be selectively reacted by a specific enzyme of E. coli.
  • the common substrate in the context of E. coli is 4-methylumbelliferyl-.beta.-D-glucuronide, which is cleaved by the E. coli typical enzyme ß-D-glucuronidase and thus releases the highly fluorescent 4-methylumbelliferone, which is measured by fluorescence optics.
  • the device according to the invention Due to the high sensitivity of the device according to the invention, which is achieved by combination of enrichment and sensitive measurement, can be dispensed with a cultivation of bacteria. Therefore, the conditions during the incubation can be tailored specifically to those of the ß-glucuronidase.
  • the volume of the reactor chamber in the measuring cell 16 is measured.
  • the signal obtained is a measure of the number of E. coli in the reactor chamber; with the known, filtered sample volume, the content of germs per unit volume can be calculated.
  • a waveform can be recorded over the time of incubation when the measuring cell or measuring device is used in the reactor chamber.
  • germs can generally be stained and measured with the modifications and combinations described below, wherein ii.) A general cell staining (eg. Vivid staining) or iii.) A selective cell staining eg with antibodies can be carried out, or also iv .) Method with the help of selective cultivation with subsequent measurement.
  • a general cell staining eg. Vivid staining
  • iii. A selective cell staining eg with antibodies can be carried out, or also iv .) Method with the help of selective cultivation with subsequent measurement.
  • the cleaning device 24 controlled by the controller 10 is moved out of the cleaning fluid vessel 25, e.g. Chlorine water, this cleaning agent, if necessary, several cleaning agents removed.
  • the sample path is flushed through as in the "Filtration" operating mode.
  • the reactor can also be rinsed with water only and backwashing of the reactor with a cleaning agent can also be carried out.
  • the measuring branch 6a can be included in the cleaning and / or backwashing. The entire cleaning process can take place automatically and is adapted to the respective prevailing conditions, such as the type of microorganisms, reagents and cleaning agents to be detected, so that after the end of the cleaning process the device is available for the measurement of a new sample
  • measuring device 38 and 39 may be arranged or provided in or for the reactor chamber 11 and the filtrate chamber 13, wherein the measuring device 38 is preferred for the reactor chamber 11 In the measuring branch 6a, the measuring device 16 downstream, further measuring devices 16a and 16b. Alternatively, these measuring devices can also be connected in parallel.
  • a seventh 3-way valve 42 between the first 3-way valve 2 and the pump 3 and the measuring branch 6b serve to circulate the measuring solution during the incubation.
  • a concentration unit 40 is inserted in front of the measuring devices 16, 16a, 16b in order to increase the sensitivity of the measurement. This can be based on chromatography, wherein via a controlled pump 41, an eluent for dissolving out the enriched analytes can be supplied.

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Claims (14)

  1. Dispositif pour surveiller périodiquement, en continu, de l'eau sur des germes microbiens, comprenant :
    un réacteur (5) ;
    au moins une conduite d'alimentation en eau (1) pour l'eau devant être analysée dans le réacteur ;
    au moins une conduite de décharge d'eau (9, 9a, 9b) sortant de la chambre de réacteur ;
    au moins une conduite d'alimentation en réactifs (18a...d) dans le réacteur et au moins un dispositif de dosage (19a...d) pour les réactifs ;
    au moins un dispositif de mesure (16) pour détecter la présence de microorganismes et/ou des produits de métabolisme de ceux-ci ;
    comprenant des moyens de pompe/de soupape (3, 4, 6, 7, 36, 42) dans les conduites d'alimentation/décharge du réacteur ;
    ainsi que comprenant un contrôleur (10) pour le contrôle programmé d'au moins les moyens de pompe/de soupape, caractérisé par le fait que
    le réacteur (5) présente une chambre de réacteur (11) et une chambre de filtrat (13) qui est séparée de cette dernière par un filtre (12), la au moins une conduite d'alimentation en eau (1) ainsi que la au moins une conduite d'alimentation en réactifs (18a...d) débouchant dans la chambre de réacteur et la au moins une conduite de décharge d'eau (9, 9a, 9b) conduisant hors de la chambre de filtrat,
    le contrôleur (10) est agencé de façon à, à l'aide des moyens de pompe/de soupape (3, 4, 6, 7), conduire une quantité d'eau pouvant être prédéfinie dans la chambre de réacteur et à travers le filtre ainsi qu'introduire une quantité de réactif pouvant être prédéfinie dans la chambre de réacteur ;
    le contrôleur (10) est agencé de façon à, à l'aide des moyens de pompe/de soupape (3, 4, 6, 7), inverser le courant d'écoulement de l'eau à travers le réacteur (5) ; et
    la chambre de filtrat (13) se trouve au-dessus de la chambre de réacteur (11).
  2. Dispositif selon la revendication 1, caractérisé par le fait que le dispositif de mesure (16) est agencé pour la mesure en aval de la chambre de filtrat (13).
  3. Dispositif selon la revendication 1, caractérisé par le fait que le dispositif de mesure (38) est agencé pour la mesure dans la chambre de réacteur (11).
  4. Dispositif selon la revendication 1, caractérisé par le fait que le dispositif de mesure (39) est agencé pour la mesure dans la chambre de filtrat (13).
  5. Dispositif selon la revendication 1, caractérisé par le fait que le contrôleur (10) est agencé de façon à, à l'aide des moyens de pompe/de soupape (3, 4, 6, 42), guider l'écoulement de courant de l'eau à travers le réacteur (5) en cercle.
  6. Dispositif selon la revendication 1, caractérisé par le fait que le contrôleur (10) est disposé pour détecter et/ou prédéfinir des volumes de mesure transportés via la pompe (3).
  7. Dispositif selon la revendication 1, caractérisé par le fait que le filtre (12) est remplaçable dans le réacteur (5).
  8. Dispositif selon la revendication 7, caractérisé par le fait qu'une plaque de changement de filtre (31) comportant au moins deux cartouches filtrantes (32a, b, c) est prévue, laquelle, rendue étanche vis-à-vis de l'enveloppe de réacteur, est déplaçable de telle sorte qu'une cartouche filtrante respective en tant que filtre actif (12) sépare les chambres de réacteur et de filtrat (11, 13).
  9. Dispositif selon la revendication 8, caractérisé par le fait qu'à la plaque de changement de filtre (31) est attribué un organe d'entraînement (33) commandé par le contrôleur (10).
  10. Dispositif selon la revendication 1, caractérisé par le filtre (12) est réalisé sensiblement plat et est incliné par rapport à l'horizontale de telle sorte que des bulles de gaz dans la chambre de réacteur (11) peuvent être dirigées vers le haut sur la surface de filtre.
  11. Dispositif selon la revendication 10, caractérisé par le fait que la chambre de filtrat (13) présente des ouvertures de conduite de décharge (9aa, 9ba) pour décharger l'eau dans une première et une deuxième conduite de décharge d'eau (9a, 9b), l'ouverture de conduite de décharge (9aa) de la première conduite de décharge d'eau (9a) étant disposée au point le plus haut de la chambre de filtrat (13) et l'ouverture de conduite de décharge (9ba) de la deuxième conduite de décharge d'eau (9b), laquelle adresse l'eau au dispositif de mesure (16), étant disposée dans un axe de symétrie (L) du réacteur (5d) qui s'étend sensiblement perpendiculairement au plan du filtre.
  12. Dispositif selon la revendication 1, caractérisé par le fait qu'au moins pour la chambre de réacteur (11), sont prévues une conduite de ventilation (34) et une soupape de ventilation (35).
  13. Dispositif selon l'une des revendications 8 et 12, caractérisé par le fait que la plaque de changement de filtre (31) présente une cavité de ventilation, qui libère une connexion entre la chambre de réacteur et/ou de filtrat (11, 13) et la conduite de ventilation conduisant vers l'extérieur dans une position de déplacement définie de la plaque.
  14. Dispositif selon la revendication 1, caractérisé par le fait qu'il présente un dispositif de nettoyage (24, 25) et que le contrôleur (10) est agencé par la commande des moyens de pompe/de soupape (2, 3, 4, 6, 7, 8) de façon à guider dans au moins un cycle de nettoyage des fluides de nettoyage à travers les conduites et unités du dispositif.
EP08756847.3A 2007-07-09 2008-07-02 Dispositif de surveillance de la présence de germes microbiens dans l'eau Active EP2165193B1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
AT0105607A AT505306B1 (de) 2007-07-09 2007-07-09 Vorrichtung zur überwachung von wasser auf mikrobielle keime
PCT/AT2008/000238 WO2009006657A1 (fr) 2007-07-09 2008-07-02 Dispositif de surveillance de la présence de germes microbiens dans l'eau

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EP2165193A1 EP2165193A1 (fr) 2010-03-24
EP2165193B1 true EP2165193B1 (fr) 2016-08-31

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US (1) US8518246B2 (fr)
EP (1) EP2165193B1 (fr)
CN (1) CN101796410B (fr)
AT (1) AT505306B1 (fr)
CA (1) CA2691827A1 (fr)
IL (1) IL203175A (fr)
RU (1) RU2478204C2 (fr)
WO (1) WO2009006657A1 (fr)

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DE102014106891A1 (de) * 2014-05-15 2015-11-19 MJM GmbH Vorrichtung zur Untersuchung von Wasserleitungen auf eine Kontamination
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WO2009006657A1 (fr) 2009-01-15
EP2165193A1 (fr) 2010-03-24
RU2010104250A (ru) 2011-08-20
US8518246B2 (en) 2013-08-27
CN101796410B (zh) 2014-05-14
AT505306A4 (de) 2008-12-15
IL203175A (en) 2012-07-31
AT505306B1 (de) 2008-12-15
CN101796410A (zh) 2010-08-04
RU2478204C2 (ru) 2013-03-27
US20100193413A1 (en) 2010-08-05

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